Endothelial cell thrombogenicity is reduced by ATRP-mediated grafting of gelatin onto PCL surfaces.

Reducing the thrombogenicity of a tissue-engineered vascular graft prior to implantation is important for improving graft patency. As functionalization of synthetic materials with cell-adhesive proteins is routinely utilized as a means to promote endothelial cell (EC) growth, we conducted detailed investigation on the proliferation and thrombogenicity of ECs on such functionalized surfaces. We observed that polycaprolactone (PCL) surfaces functionalized with poly(glycidyl methacrylate) [(P(GMA)] brushes via atom transfer radical polymerization (ATRP) alone resulted in the enhancement of an activated EC profile characterized by low production of nitric oxide (NO), platelet activation and elevated expression levels of von Willebrand factor (vWF) and matrix metalloproteinase-2 (MMP-2). When gelatin was conjugated onto the PCL-g-P(GMA) surfaces, not only were EC proliferation and endothelial coverage significantly improved, but an anti-thrombogenic profile was also observed. We demonstrated that PCL can be successfully functionalized by a controllable surface-initiated polymerization method and importantly, the thrombogenic profile of the endothelial cells can be influenced by material surface chemistry (e.g. the presence of polymer graft chains). Our findings emphasize the importance of a careful consideration of materials for vascular graft applications, as well as differential endothelial cell physiology on surfaces with different material chemistry.

[1]  Changyou Gao,et al.  Surface modification of polycaprolactone membrane via aminolysis and biomacromolecule immobilization for promoting cytocompatibility of human endothelial cells. , 2002, Biomacromolecules.

[2]  D. Cheek,et al.  Nitric oxide and regulation of vascular tone: pharmacological and physiological considerations. , 1998, American journal of critical care : an official publication, American Association of Critical-Care Nurses.

[3]  C. Choong,et al.  Multifunctional P(PEGMA)-REDV conjugated titanium surfaces for improved endothelial cell selectivity and hemocompatibility. , 2013, Journal of materials chemistry. B.

[4]  W. Yuan,et al.  PCL film surfaces conjugated with P(DMAEMA)/gelatin complexes for improving cell immobilization and gene transfection. , 2011, Bioconjugate chemistry.

[5]  Gregory Y H Lip,et al.  The adhesion molecule P-selectin and cardiovascular disease. , 2003, European heart journal.

[6]  F. Huang,et al.  Proinflammatory activation of macrophages by bisphenol A-glycidyl-methacrylate involved NFκB activation via PI3K/Akt pathway. , 2012, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[7]  J. Coltart,et al.  ‘Miracle stents’ - a future without restenosis , 2007, McGill journal of medicine : MJM : an international forum for the advancement of medical sciences by students.

[8]  F. D. de Wolf,et al.  Secreted production of a custom-designed, highly hydrophilic gelatin in Pichia pastoris. , 2001, Protein engineering.

[9]  Dietmar W Hutmacher,et al.  Co-culture of bone marrow fibroblasts and endothelial cells on modified polycaprolactone substrates for enhanced potentials in bone tissue engineering. , 2006, Tissue engineering.

[10]  S. Watson,et al.  Platelet-collagen interaction: is GPVI the central receptor? , 2003, Blood.

[11]  Steven G Wise,et al.  A multilayered synthetic human elastin/polycaprolactone hybrid vascular graft with tailored mechanical properties. , 2011, Acta biomaterialia.

[12]  R. H. Sohn,et al.  Regulation of endothelial thrombomodulin expression by inflammatory cytokines is mediated by activation of nuclear factor-kappa B. , 2005, Blood.

[13]  C. Choong,et al.  Simple surface modification of poly(ε-caprolactone) for apatite deposition from simulated body fluid , 2005 .

[14]  S. Teoh,et al.  Processing methods of ultrathin poly(epsilon-caprolactone) films for tissue engineering applications. , 2007, Biomacromolecules.

[15]  R. Cannon,et al.  Interactions between nitric oxide and endothelin in the regulation of vascular tone of human resistance vessels in vivo. , 2000, Hypertension.

[16]  C. Satriano,et al.  The effect of irradiation modification and RGD sequence adsorption on the response of human osteoblasts to polycaprolactone. , 2005, Biomaterials.

[17]  K. Neoh,et al.  Antibacterial inorganic-organic hybrid coatings on stainless steel via consecutive surface-initiated atom transfer radical polymerization for biocorrosion prevention. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[18]  J. Feijen,et al.  Relation between cell density and the secretion of von Willebrand factor and prostacyclin by human umbilical vein endothelial cells. , 2001, Biomaterials.

[19]  H. Bitterman,et al.  Regulation of Endothelial Matrix Metalloproteinase-2 by Hypoxia/Reoxygenation , 2002, Circulation research.

[20]  Robert Gurny,et al.  Degradation and Healing Characteristics of Small-Diameter Poly(&egr;-Caprolactone) Vascular Grafts in the Rat Systemic Arterial Circulation , 2008, Circulation.

[21]  F. Huang,et al.  The upregulation of tumour necrosis factor-α and surface antigens expression on macrophages by bisphenol A-glycidyl-methacrylate. , 2012, International endodontic journal.

[22]  Alison P McGuigan,et al.  The influence of biomaterials on endothelial cell thrombogenicity. , 2007, Biomaterials.

[23]  Christopher Breuer,et al.  Artificial blood vessel: the Holy Grail of peripheral vascular surgery. , 2005, Journal of vascular surgery.

[24]  E. Jaffe,et al.  Synthesis of von Willebrand factor by cultured human endothelial cells. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[25]  I. Elalamy,et al.  Platelet release of trimolecular complex components MT1-MMP/TIMP2/MMP2: involvement in MMP2 activation and platelet aggregation. , 2000, Blood.

[26]  Maureane Hoffman,et al.  Remodeling the Blood Coagulation Cascade , 2003, Journal of Thrombosis and Thrombolysis.

[27]  D. Yan,et al.  Surface modification of polycaprolactone membrane via layer-by-layer deposition for promoting blood compatibility. , 2008, Journal of biomedical materials research. Part B, Applied biomaterials.

[28]  C. Choong,et al.  Surface modification of polycaprolactone substrates using collagen-conjugated poly(methacrylic acid) brushes for the regulation of cell proliferation and endothelialisation , 2012 .

[29]  B. Lämmle,et al.  von Willebrand factor-mediated platelet adhesion is critical for deep vein thrombosis in mouse models. , 2011, Blood.

[30]  D. Collen,et al.  Thrombomodulin-protein C-EPCR system: integrated to regulate coagulation and inflammation. , 2004, Arteriosclerosis, thrombosis, and vascular biology.

[31]  S. Moncada,et al.  ENDOGENOUS NITRIC OXIDE INHIBITS HUMAN PLATELET ADHESION TO VASCULAR ENDOTHELIUM , 1987, The Lancet.

[32]  F. Xu,et al.  Surface functionalization of polycaprolactone films via surface-initiated atom transfer radical polymerization for covalently coupling cell-adhesive biomolecules. , 2010, Biomaterials.

[33]  L. Ghasemi‐Mobarakeh,et al.  Electrospun poly(epsilon-caprolactone)/gelatin nanofibrous scaffolds for nerve tissue engineering. , 2008, Biomaterials.

[34]  D. Hutmacher,et al.  The return of a forgotten polymer : Polycaprolactone in the 21st century , 2009 .

[35]  R. Bach Initiation of Coagulation by Tissue Facto , 1988 .

[36]  J. Sixma,et al.  Platelet adhesion to collagen types I through VIII under conditions of stasis and flow is mediated by GPIa/IIa (alpha 2 beta 1-integrin). , 1994, Blood.

[37]  M. Shiotani,et al.  Graft polymerization of methacrylic acid onto polytetrafluoroethylene initiated by alkyllithium/electron‐donating solvents , 2004 .

[38]  S. Teoh,et al.  Scaffold design and in vitro study of osteochondral coculture in a three-dimensional porous polycaprolactone scaffold fabricated by fused deposition modeling. , 2003, Tissue engineering.

[39]  B. Jacobson,et al.  Extracellular calcium regulates HeLa cell morphology during adhesion to gelatin: role of translocation and phosphorylation of cytosolic phospholipase A2. , 1998, Molecular biology of the cell.

[40]  Nicolas H Voelcker,et al.  Stimuli-responsive interfaces and systems for the control of protein-surface and cell-surface interactions. , 2009, Biomaterials.

[41]  T. Mihaljevic,et al.  Superior late patency of small-diameter Dacron grafts seeded with omental microvascular cells: an experimental study. , 1994, The Annals of thoracic surgery.

[42]  F. W. Blaisdell,et al.  Why small caliber vascular grafts fail: a review of clinical and experimental experience and the significance of the interaction of blood at the interface. , 1986, The Journal of surgical research.

[43]  U. Pendurthi,et al.  Tissue factor-factor VIIa signaling. , 2004, Arteriosclerosis, thrombosis, and vascular biology.

[44]  T. Wakefield,et al.  Macrovascular thrombosis is driven by tissue factor derived primarily from the blood vessel wall. , 2005, Blood.

[45]  R. Kempczinski,et al.  Impact of endothelial cell seeding on long-term patency and subendothelial proliferation in a small-caliber highly porous polytetrafluoroethylene graft. , 1987, Journal of vascular surgery.

[46]  A. McGuigan,et al.  Tissue factor and thrombomodulin expression on endothelial cell-seeded collagen modules for tissue engineering. , 2007, Journal of biomedical materials research. Part A.

[47]  R. Virmani,et al.  Pathological Correlates of Late Drug-Eluting Stent Thrombosis: Strut Coverage as a Marker of Endothelialization , 2007, Circulation.